海产品中副溶血弧菌变性高效液相色谱检测技术研究

Determination of Vibrio Parahaemolyticus in Seafood by Denaturing High Performance Liquid Chromatography

目的建立快速、准确检测海产品中副溶血弧菌的新方法。方法应用聚合酶链式反应(PCR)结合变性高效液相色谱(DHPLC)技术对海产品中致病菌(副溶血弧菌)进行检测,探讨其特异性和灵敏度。利用DHPLC在非变性温度下进行双链DNA分离的原理及DHPLC技术分析副溶血弧菌特异性PCR扩增产物。在优化的分析条件下,对样品洗脱峰排列形成的聚类分析图进行分析,得到副溶血弧菌的特征峰型图谱。结果该方法有很好的特异性,与传统检测方法进行比较其准确度为100%。结论本实验建立的海产品中副溶血弧菌PCR-DHPLC检测技术,特异性、灵敏度、准确度高,操作快速简便,在海产品安全检测中具有重要应用价值。

Objective To detect Vibrio parahaemolyticus in seafood by PCR with denaturing high performance liquid chromatography（DHPLC） and explore its sensitivity and specificity of the new method for Vibrio parahaemolyticus in seafood.Methods DHPLC was used for seperating double-stranded DNA under non-degeneration temperature and for detecting the specific PCR amplification products.Under the optimum conditions,the clustering analysis diagram of elution peaks of samples were analyzed and the spectrum with characteristic peak of Vibrio parahaemolyticus was obtained.Results The method had excellent specificity and accuracy,and the method accuracy was 100%,Compared with the traditional detection methods.Conclusion PCR-DHPLC detection method for Vibrio parahaemolyticus in seafood has good specificity,sensitivity,high accuracy and rapid operation and simplicity. It has an important application value in the seafood safety inspection.